Synthesis and characteristics of the human serum albumin-triazine chiral stationary phase

Human serum albumin (HSA) was successfully bonded to silica with s‐triazine as activator. The coupling reaction by this method was rapid and effective. The triazine‐activated silica is relatively stable and can be installed for at least 1 month without obvious loss of reactivity when stored below 30°C, pH below 7. It was observed that the amount of bound HSA reached 120 mg/g silica calculated from the UV absorbance difference of the HSA solution. d,l‐tryptophan was selected as the probe solute to characterize the properties of HSA bonded s‐triazine chiral stationary phase, and separation factor of 9.4 was obtained for d,l‐tryptophan. Furthermore, the amount of effective HSA on silica was measured by high‐performance frontal analysis, and only 16.8 mg/g silica was responsible for the resolution of d,l‐tryptophan. These results indicate that the amount of both the bound and effective HSA on silica with triazine as activator was much higher than those by the Schiff base coupling method. Different kinds of enantiomers were resolved successfully on the aminopropylsilica‐bonded HSA s‐triazine chiral stationary phase. Chirality 12:714–719, 2000. © 2000 Wiley‐Liss, Inc.